Burst release type artificial bear gall powder nano preparation as well as preparation method and application of burst release type artificial bear gall powder nano preparation

Abstract

The invention discloses a burst release type artificial bear gall powder nano preparation which has the characteristics of high biocompatibility and biological activity, pH response type burst release and high drug loading capacity and stability, and also discloses a preparation method of the burst release type artificial bear gall powder nano preparation. Comprising the following steps: preparing magnesium ion-doped calcium phosphate nanoparticles, loading artificial bear gall powder, collecting and purifying, and freeze-drying. The method has the characteristics of simplicity in operation, mild conditions and easiness in large-scale production. The nano preparation can be widely applied to preparation of drugs for treating diseases needing rapid release of the drugs in an acidic microenvironment.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical formulation technology, specifically to a burst-release artificial bear bile powder nanoparticle formulation, its preparation method, and its application. Background Technology

[0002] Artificial bear bile powder (ABP), an important traditional Chinese medicine, possesses a variety of pharmacological activities, including clearing heat and detoxifying, calming the liver and improving eyesight, promoting bile secretion and dissolving stones, and anti-inflammatory and analgesic effects. Therefore, it is widely used clinically to treat hepatobiliary diseases, ophthalmological conditions, cardiovascular and cerebrovascular diseases, and various inflammatory diseases. However, despite its significant pharmacological effects, artificial bear bile powder still faces many challenges in actual oral administration. Its main active ingredients, such as ursodeoxycholic acid, often suffer from low bioavailability, insufficient chemical stability, easy degradation in the acidic environment of the stomach, and non-specific distribution, making it difficult to effectively accumulate in target tissues or organs. These factors collectively limit its full clinical efficacy and further expansion of its application scope.

[0003] Nanoparticle drug delivery systems have attracted much attention in recent years due to their unique advantages. They can significantly improve the solubility and stability of poorly soluble drugs, enhance drug bioavailability, and achieve precise targeted delivery and controllable drug release, making them a cutting-edge strategy for addressing the challenges of traditional drug delivery. Among various carrier materials, calcium phosphate materials have been widely explored as drug delivery carriers due to their excellent biocompatibility, good biodegradability, and superior osteointegration capabilities. However, calcium phosphate carrier materials prepared by traditional methods often have limitations in structural regulation and functional design, lacking sufficient pH response precision. This makes it difficult to achieve rapid, efficient, and on-demand drug release under complex physiological environments, thus limiting their potential for clinical application to some extent. Summary of the Invention

[0004] To address the technical problems existing in the prior art, this invention provides a burst-release artificial bear bile powder nanoparticle formulation, its preparation method, and its application. The method involves doping calcium phosphate nanoparticles with magnesium ions (Mg²⁺) and using anhydrous ethanol as a loading solvent to prepare an artificial bear bile powder nanoparticle formulation with good bioactivity and pH-based burst-release characteristics. Magnesium ion doping can alter the crystal structure and surface properties of calcium phosphate nanoparticles, further enhancing their bioactivity and enabling them to exhibit more significant dissolution and burst-release behavior in acidic environments. This holds promise for achieving targeted and rapid release of artificial bear bile powder from specific pathological sites (such as inflammation or tumor microenvironments) or the intestines, improving therapeutic efficacy and reducing side effects.

[0005] To achieve the above objectives, the present invention provides the following technical solution: A method for preparing a burst-release artificial bear bile powder nanoparticle formulation includes the following steps: Step S1: Preparation of magnesium ion-doped calcium phosphate nanoparticles Step S1.1: Dissolve calcium salt and magnesium salt in deionized water to obtain a mixed cation solution, wherein the molar ratio of magnesium ions to calcium ions is 0.05:1 to 0.2:1; Step S1.2: Dissolve phosphate in deionized water to obtain a phosphate solution, wherein the molar ratio of phosphate ions to calcium ions is 1.6:1 to 1.8:1; Step S1.3: Under continuous stirring at 100-500 rpm using a magnetic stirrer, the phosphate solution is added dropwise to the mixed cation solution, while the pH of the mixture is adjusted to 9.0-11.0. The mixture is reacted at 60-80℃ for 8-12 hours to obtain magnesium-doped calcium phosphate precipitate. Step S1.4: Remove impurities from the precipitate and dry it to obtain magnesium ion-doped calcium phosphate nanoparticles. Step S2, Loading with artificial bear bile powder Step S2.1: Disperse the magnesium ion-doped calcium phosphate nanoparticles obtained in step S1.4 in anhydrous ethanol to obtain a nanoparticle dispersion. Step S2.2: Dissolve the artificial bear bile powder in anhydrous ethanol to obtain an ethanol solution or dispersion of the artificial bear bile powder; Step S2.3: Add the ethanol solution or dispersion of the artificial bear bile powder obtained in step S2.2 to the nanoparticle dispersion obtained in step S2.1 and stir continuously. The mass ratio of magnesium ion-doped calcium phosphate nanoparticles to artificial bear bile powder is 4:1 to 6:1 to obtain a mixed suspension. Step S2.4: The mixed suspension obtained in step S2.3 is subjected to ultrasonic-assisted light-protected vibration at a temperature of 25-35℃ for 12-24 hours to complete the drug loading and obtain the drug-loaded suspension. Step S3: Collection and Purification Step S3.1: Centrifuge the drug-loaded suspension obtained in step S2.4 and collect the precipitate; Step S3.2: Wash the precipitate obtained in step S3.1 with anhydrous ethanol; Step S4: Using freeze-drying, the precipitate obtained in step S3.2 is pre-frozen at -40℃ to -80℃ for 2-12 hours, and then sublimated and dried under a vacuum pressure of 10-30 Pa for 16-48 hours to obtain a burst-release artificial bear bile powder nanoparticle formulation.

[0006] Furthermore, in step S1.1, the calcium salt and magnesium salt need to be completely dissolved in deionized water by magnetic stirring at a temperature of 15-30℃. The calcium salt is calcium chloride dihydrate and the magnesium salt is magnesium chloride. In step S1.2, the phosphate needs to be completely dissolved in deionized water by magnetic stirring at a temperature of 15-30℃. The phosphate is diammonium hydrogen phosphate. In step S1.3, the pH adjustment process of the mixture is to add alkaline pH adjuster ammonia water dropwise by a metering pump connected to an automatic pH controller, and monitor the pH of the mixture in real time so that the pH value is dynamically maintained in the range of 9.0-11.0.

[0007] Furthermore, in step S2.1, the process of dispersing magnesium ion-doped calcium phosphate nanoparticles in anhydrous ethanol is specifically as follows: a high-shear disperser is used to pre-disperse the nanoparticles at a speed of 3000 rpm for 10 min, and then a high-pressure homogenizer is used to homogenize the nanoparticles five times at a pressure of 150 MPa to obtain a uniformly dispersed nanoparticle dispersion. In step S2.2, the particle size requirement for artificial bear bile powder is D90 < 50 µm.

[0008] Further, step S3.1 specifically involves centrifuging the drug-loaded suspension obtained in step S2.4 at a speed of 1000-4000 rpm for 10-30 min, then collecting the precipitate, and repeatedly centrifuging and redispersing the collected precipitate with anhydrous ethanol, washing it 2-5 times, and collecting the precipitate again.

[0009] A burst-release artificial bear bile powder nanoformulation prepared by the above method.

[0010] Furthermore, the cumulative release rate of the main active ingredients in the artificial bear bile powder was less than 40% within 24 hours at 37°C in a phosphate buffer solution with pH 7.4; and greater than 40% within 2 hours and greater than 70% within 24 hours in a phosphate buffer solution with pH 5.5 at 37°C.

[0011] Furthermore, the drug loading of the formulation is 10-20%.

[0012] Furthermore, the formulation is a composite nanoparticle powder, the physical structure of which is that artificial bear bile powder is loaded on a porous framework formed by the self-assembly of magnesium ion-doped calcium phosphate nanocrystals. The framework is composed of nanoparticle aggregates, a network of interlaced nanosheets / nanose needles, or a flower-like structure of micron-scale sheet stacks.

[0013] Furthermore, it includes artificial bear bile powder and zinc oxide nanoparticles, wherein the artificial bear bile powder comprises the following components: Tauroursodeoxycholic acid 40-55%; Taurine chenodeoxycholic acid 15-30%; Ursodeoxycholic acid 2-5%; Chenodeoxycholic acid 1-3%; Bile acids 5-10%; The bile acid components satisfy the following proportional relationship, specifically: a) The mass ratio of tauroursodeoxycholic acid (TUDCA) to taurourcheideoxycholic acid (TCDCA) is (1.5-2.5):1; b) The mass fraction of taurine-conjugated bile acids in total bile acids is greater than 80% and less than or equal to 100%, and the taurine-conjugated bile acids are the sum of tauroursodeoxycholic acid (TUDCA) and taurourchedeoxycholic acid (TCDCA). Total bile acids include free bile acids, glycine-conjugated bile acids, and taurine-conjugated bile acids.

[0014] The above-mentioned burst-release artificial bear bile powder nanoparticles are used in the preparation of drugs for treating diseases that require rapid drug release in an acidic microenvironment, with the diseases selected from hepatobiliary inflammation or inflammatory bowel disease.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows: (1) The burst-release artificial bear bile powder nano-formulation provided by the present invention has high biocompatibility and bioactivity. Calcium phosphate is selected as the carrier, and its excellent biocompatibility is utilized. By doping with bioactive magnesium ions, the bioactivity of the nano-carrier is further enhanced. In synergy with artificial bear bile powder, it can promote its better pharmacological effects.

[0016] (2) The burst-release artificial bear bile powder nano-formulation provided by the present invention has the characteristics of pH-responsive burst release. The doping of magnesium ions significantly improves the dissolution rate of calcium phosphate nanoparticles in acidic environment. Under acidic conditions (pH 5.5) that simulate the gastrointestinal tract, inflammation or tumor microenvironment, the drug can achieve rapid burst release, while under neutral environment (pH 7.4) it maintains slow release, thus achieving precise release and targeted delivery of the drug.

[0017] (3) The burst-release artificial bear bile powder nano-formulation provided by the present invention has high drug loading and stability. The nano-formulation prepared by the preparation method disclosed in the present invention has a large specific surface area and pore structure. Combined with the optimized loading process, it can achieve a high drug loading of artificial bear bile powder, ensure the effective dose for treatment, and help improve the stability and bioavailability of artificial bear bile powder.

[0018] (4) The method for preparing burst-release artificial bear bile powder nanoparticles provided by the present invention has the characteristics of simple operation, mild conditions, and easy scale-up production, laying the foundation for the industrial production of artificial bear bile powder nanoparticles.

[0019] (5) The burst-release artificial bear bile powder nano-formulation provided by the present invention can improve drug utilization and reduce side effects based on its own effects. Specifically, it can reduce drug accumulation in non-target sites through targeted and controlled release, thereby improving the therapeutic effect of artificial bear bile powder and reducing systemic side effects. Therefore, it can be widely used in the preparation of drugs for treating diseases that require rapid release of drugs in an acidic microenvironment. The diseases are selected from hepatobiliary inflammation or inflammatory bowel disease. Attached Figure Description

[0020] The disclosure of this invention is illustrated with reference to the accompanying drawings. It should be understood that the drawings are for illustrative purposes only and are not intended to limit the scope of protection of this invention. In the drawings, the same reference numerals are used to refer to the same parts. Wherein: Figure 1 These are XRD patterns of the burst-release artificial bear bile powder nanoformulations prepared in Examples 1-3; Figure 2 Here is a SEM image of the burst-release artificial bear bile powder nanoformulation prepared in Example 1; Figure 3 Here is a SEM image of the burst-release artificial bear bile powder nanoformulation prepared in Example 2; Figure 4 This is a SEM image of the burst-release artificial bear bile powder nanoformulation prepared in Example 3. Detailed Implementation

[0021] It is readily understood that, based on the technical solution of this invention, those skilled in the art can propose various interchangeable structural methods and implementations without altering the essential spirit of the invention. Therefore, the following detailed embodiments and accompanying drawings are merely illustrative examples of the technical solution of this invention and should not be considered as the entirety of the invention or as limitations or restrictions on the technical solution of this invention.

[0022] The technical solution of this application will now be described in detail with reference to the accompanying drawings. The description of exemplary embodiments is merely for illustrative purposes and is by no means a limitation on the invention or its application or use.

[0023] Example 1 A method for preparing a burst-release artificial bear bile powder nanoparticle formulation includes the following steps: Step S1: Preparation of magnesium ion-doped calcium phosphate nanoparticles Step S1.1: Dissolve calcium salt and magnesium salt in deionized water to obtain a mixed cation solution, wherein the molar ratio of magnesium ions to calcium ions is 0.05:1; It should be noted that in this embodiment, the calcium salt and magnesium salt need to be completely dissolved in deionized water by magnetic stirring at a temperature of 15°C. The calcium salt is calcium chloride dihydrate and the magnesium salt is magnesium chloride. Step S1.2: Dissolve phosphate in deionized water to obtain a phosphate solution, wherein the molar ratio of phosphate ions to calcium ions is 1.6:1; It should be noted that in this embodiment, the phosphate needs to be completely dissolved in deionized water by magnetic stirring at a temperature of 15°C, and the phosphate is diammonium hydrogen phosphate. Step S1.3: Under continuous stirring at 100 rpm using a magnetic stirrer, the phosphate solution is added dropwise to the mixed cation solution, while the pH of the mixture is adjusted to 9.0. The mixture is reacted at 60°C for 8 hours to obtain magnesium-doped calcium phosphate precipitate. It should be noted that in this embodiment, the pH adjustment process of the mixture is to add alkaline pH adjuster ammonia water dropwise by a metering pump connected to an automatic pH controller, and monitor the pH of the mixture in real time so that the pH is dynamically maintained within the required range. Step S1.4: Remove impurities from the precipitate and dry it to obtain magnesium ion-doped calcium phosphate nanoparticles. Step S2, Loading with artificial bear bile powder Step S2.1: Disperse the magnesium ion-doped calcium phosphate nanoparticles obtained in step S1.4 in anhydrous ethanol to obtain a nanoparticle dispersion. It should be noted that in this embodiment, the process of dispersing magnesium ion-doped calcium phosphate nanoparticles in anhydrous ethanol is as follows: a high shear disperser is used to pre-disperse the nanoparticles at a speed of 3000 rpm for 10 min, and then a high pressure homogenizer is used to homogenize the nanoparticles five times at a pressure of 150 MPa to obtain a uniformly dispersed nanoparticle dispersion. Step S2.2: Dissolve the artificial bear bile powder in anhydrous ethanol to obtain an ethanol solution or dispersion of the artificial bear bile powder; it should be noted that the particle size requirement of the artificial bear bile powder is D90 < 50 µm. Step S2.3: Add the ethanol solution or dispersion of the artificial bear bile powder obtained in step S2.2 to the nanoparticle dispersion obtained in step S2.1 and stir continuously. The mass ratio of magnesium ion-doped calcium phosphate nanoparticles to artificial bear bile powder is 6:1 to obtain a mixed suspension. Step S2.4: The mixed suspension obtained in step S2.3 is subjected to ultrasonic-assisted light-protected vibration at 25°C for 12 hours to complete the drug loading and obtain the drug-loaded suspension. Step S3: Collection and Purification Step S3.1: Centrifuge the drug-loaded suspension obtained in step S2.4 at 1000 rpm for 10 min, then collect the precipitate, and repeat the centrifugation and redispersion of the collected precipitate with anhydrous ethanol, wash twice, and collect the precipitate again. Step S3.2: Wash the precipitate obtained in step S3.1 with anhydrous ethanol; Step S4: Using freeze-drying, the precipitate obtained in step S3.2 is pre-frozen at -40℃ for 12 hours, and then sublimated and dried under a vacuum pressure of 30 Pa for 16 hours to obtain a burst-release artificial bear bile powder nanoparticle formulation. Figure 1 The XRD pattern of the burst-release artificial bear bile powder nanoparticle formulation is shown. The phase retrieval results indicate that the material was successfully prepared.

[0024] This embodiment uses the above method to prepare a burst-release artificial bear bile powder nanoformulation: (1) The cumulative release rate of the main active ingredients in artificial bear bile powder in phosphate buffer solution at pH 7.4 at 37°C for 24 hours was 38%; the cumulative release rate of the main active ingredients in artificial bear bile powder in phosphate buffer solution at pH 5.5 at 37°C for 2 hours was 40.5%; and the cumulative release rate of the main active ingredients in artificial bear bile powder in 24 hours was 75%.

[0025] (2) Its drug loading is 14.5%.

[0026] (3) It is a composite nanoparticle powder with the following physical structure, such as Figure 2 As shown, artificial bear bile powder is loaded onto a porous framework formed by the self-assembly of magnesium ion-doped calcium phosphate nanocrystals. The framework consists of nanoparticle aggregates, a network of interlaced nanosheets / nanose needles, or a flower-like structure of stacked micron-scale sheets.

[0027] (4) Its components include artificial bear bile powder and zinc oxide nanoparticles, and the artificial bear bile powder includes the following components: Tauroursodeoxycholic acid 40-55%; Taurine chenodeoxycholic acid 15-30%; Ursodeoxycholic acid 2-5%; Chenodeoxycholic acid 1-3%; Bile acids 5-10%; The bile acid components satisfy the following ratio: the mass ratio of tauroursodeoxycholic acid (TUDCA) to taurourcheideoxycholic acid (TCDCA) is 1.5:1.

[0028] The above-mentioned burst-release artificial bear bile powder nanoparticles are used in the preparation of drugs for treating diseases that require rapid drug release in an acidic microenvironment, with the diseases being selected from hepatobiliary inflammatory diseases.

[0029] In vitro release: Example

[0030] A method for preparing a burst-release artificial bear bile powder nanoparticle formulation includes the following steps: Step S1: Preparation of magnesium ion-doped calcium phosphate nanoparticles Step S1.1: Dissolve calcium salt and magnesium salt in deionized water to obtain a mixed cation solution, wherein the molar ratio of magnesium ions to calcium ions is 0.1:1; It should be noted that in this embodiment, the calcium salt and magnesium salt need to be completely dissolved in deionized water by magnetic stirring at a temperature of 25°C. The calcium salt is calcium chloride dihydrate and the magnesium salt is magnesium chloride. Step S1.2: Dissolve phosphate in deionized water to obtain a phosphate solution, wherein the molar ratio of phosphate ions to calcium ions is 1.7:1. It should be noted that in this embodiment, the phosphate needs to be completely dissolved in deionized water by magnetic stirring at a temperature of 25°C, and the phosphate is diammonium hydrogen phosphate. Step S1.3: Under continuous stirring at 500 rpm using a magnetic stirrer, the phosphate solution is added dropwise to the mixed cation solution, while the pH of the mixture is adjusted to 10. The mixture is reacted at 70°C for 10 hours to obtain magnesium-doped calcium phosphate precipitate. It should be noted that in this embodiment, the pH adjustment process of the mixture is to add alkaline pH adjuster ammonia water dropwise by a metering pump connected to an automatic pH controller, and monitor the pH of the mixture in real time so that the pH is dynamically maintained within the required range. Step S1.4: Remove impurities from the precipitate and dry it to obtain magnesium ion-doped calcium phosphate nanoparticles. Step S2, Loading with artificial bear bile powder Step S2.1: Disperse the magnesium ion-doped calcium phosphate nanoparticles obtained in step S1.4 in anhydrous ethanol to obtain a nanoparticle dispersion. It should be noted that in this embodiment, the process of dispersing magnesium ion-doped calcium phosphate nanoparticles in anhydrous ethanol is as follows: a high shear disperser is used to pre-disperse the nanoparticles at a speed of 3000 rpm for 10 min, and then a high pressure homogenizer is used to homogenize the nanoparticles five times at a pressure of 150 MPa to obtain a uniformly dispersed nanoparticle dispersion. Step S2.2: Dissolve the artificial bear bile powder in anhydrous ethanol to obtain an ethanol solution or dispersion of the artificial bear bile powder; it should be noted that the particle size requirement of the artificial bear bile powder is D90 < 50 µm. Step S2.3: Add the ethanol solution or dispersion of the artificial bear bile powder obtained in step S2.2 to the nanoparticle dispersion obtained in step S2.1 and stir continuously. The mass ratio of magnesium ion-doped calcium phosphate nanoparticles to artificial bear bile powder is 5:1 to obtain a mixed suspension. Step S2.4: The mixed suspension obtained in step S2.3 is subjected to ultrasonic-assisted light-protected vibration at 30°C for 18 hours to complete the drug loading and obtain the drug-loaded suspension. Step S3: Collection and Purification Step S3.1: Centrifuge the drug-loaded suspension obtained in step S2.4 at 2500 rpm for 20 min, then collect the precipitate, and repeat the centrifugation and redispersion of the collected precipitate with anhydrous ethanol, and wash it 3 times, and collect the precipitate again. Step S3.2: Wash the precipitate obtained in step S3.1 with anhydrous ethanol; Step S4: Using freeze-drying, the precipitate obtained in step S3.2 is pre-frozen at -60℃ for 7 hours, and then sublimated and dried under a vacuum pressure of 15 Pa for 32 hours to obtain a burst-release artificial bear bile powder nanoparticle formulation. Figure 1 The XRD pattern of the burst-release artificial bear bile powder nanoparticle formulation is shown. The phase retrieval results indicate that the material was successfully prepared.

[0031] This embodiment uses the above method to prepare a burst-release artificial bear bile powder nanoformulation: (1) The cumulative release rate of the main active ingredients in artificial bear bile powder in phosphate buffer solution at pH 7.4 at 37°C for 24 hours was 38%; the cumulative release rate of the main active ingredients in artificial bear bile powder in phosphate buffer solution at pH 5.5 at 37°C for 2 hours was 42.1%; and the cumulative release rate of the main active ingredients in artificial bear bile powder in 24 hours was 82.3%.

[0032] (2) Its drug loading is 16.5%.

[0033] (3) It is a composite nanoparticle powder with the following physical structure, such as Figure 3 As shown, artificial bear bile powder is loaded onto a porous framework formed by the self-assembly of magnesium ion-doped calcium phosphate nanocrystals. The framework consists of nanoparticle aggregates, a network of interlaced nanosheets / nanose needles, or a flower-like structure of stacked micron-scale sheets.

[0034] (4) Its components include artificial bear bile powder and zinc oxide nanoparticles, and the artificial bear bile powder includes the following components: Tauroursodeoxycholic acid 40-55%; Taurine chenodeoxycholic acid 15-30%; Ursodeoxycholic acid 2-5%; Chenodeoxycholic acid 1-3%; Bile acids 5-10%; The bile acid components satisfy the following proportional relationship: The mass fraction of taurine-conjugated bile acids in total bile acids is greater than 80% and less than or equal to 100%, and the taurine-conjugated bile acids are the sum of tauroursodeoxycholic acid (TUDCA) and taurourchedeoxycholic acid (TCDCA). Total bile acids include free bile acids, glycine-conjugated bile acids, and taurine-conjugated bile acids.

[0035] The above-mentioned burst-release artificial bear bile powder nanoparticle formulation is used in the preparation of drugs for treating diseases that require rapid drug release in an acidic microenvironment, with the disease being inflammatory bowel disease.

[0036] Example 3 A method for preparing a burst-release artificial bear bile powder nanoparticle formulation includes the following steps: Step S1: Preparation of magnesium ion-doped calcium phosphate nanoparticles Step S1.1: Dissolve calcium and magnesium salts in deionized water to obtain a mixed cation solution, wherein the molar ratio of magnesium ions to calcium ions is 0.2:1; It should be noted that in this embodiment, the calcium salt and magnesium salt need to be completely dissolved in deionized water by magnetic stirring at a temperature of 30°C. The calcium salt is calcium chloride dihydrate and the magnesium salt is magnesium chloride. Step S1.2: Dissolve phosphate in deionized water to obtain a phosphate solution, wherein the molar ratio of phosphate ions to calcium ions is 1.8:1; It should be noted that in this embodiment, the phosphate needs to be completely dissolved in deionized water by magnetic stirring at a temperature of 30°C, and the phosphate is diammonium hydrogen phosphate. Step S1.3: Under continuous stirring at 500 rpm using a magnetic stirrer, the phosphate solution is added dropwise to the mixed cation solution, while the pH of the mixture is adjusted to 11.0. The mixture is reacted at 80°C for 12 hours to obtain magnesium-doped calcium phosphate precipitate. It should be noted that in this embodiment, the pH adjustment process of the mixture is to add alkaline pH adjuster ammonia water dropwise by a metering pump connected to an automatic pH controller, and monitor the pH of the mixture in real time so that the pH is dynamically maintained within the required range. Step S1.4: Remove impurities from the precipitate and dry it to obtain magnesium ion-doped calcium phosphate nanoparticles. Step S2, Loading with artificial bear bile powder Step S2.1: Disperse the magnesium ion-doped calcium phosphate nanoparticles obtained in step S1.4 in anhydrous ethanol to obtain a nanoparticle dispersion. It should be noted that in this embodiment, the process of dispersing magnesium ion-doped calcium phosphate nanoparticles in anhydrous ethanol is as follows: a high shear disperser is used to pre-disperse the nanoparticles at a speed of 3000 rpm for 10 min, and then a high pressure homogenizer is used to homogenize the nanoparticles five times at a pressure of 150 MPa to obtain a uniformly dispersed nanoparticle dispersion. Step S2.2: Dissolve the artificial bear bile powder in anhydrous ethanol to obtain an ethanol solution or dispersion of the artificial bear bile powder; it should be noted that the particle size requirement of the artificial bear bile powder is D90 < 50 µm. Step S2.3: Add the ethanol solution or dispersion of the artificial bear bile powder obtained in step S2.2 to the nanoparticle dispersion obtained in step S2.1 and stir continuously. The mass ratio of magnesium ion-doped calcium phosphate nanoparticles to artificial bear bile powder is 4:1 to obtain a mixed suspension. Step S2.4: The mixed suspension obtained in step S2.3 is subjected to ultrasonic-assisted light-protected vibration at 35°C for 24 hours to complete the drug loading, resulting in a drug-loaded suspension. Step S3: Collection and Purification Step S3.1: Centrifuge the drug-loaded suspension obtained in step S2.4 at 4000 rpm for 30 min, then collect the precipitate, and repeat the centrifugation and redispersion of the collected precipitate with anhydrous ethanol, and wash it 5 times, and collect the precipitate again. Step S3.2: Wash the precipitate obtained in step S3.1 with anhydrous ethanol; Step S4: Using freeze-drying, the precipitate obtained in step S3.2 is pre-frozen at -80℃ for 2 hours, and then sublimated and dried under a vacuum pressure of 10 Pa for 48 hours to obtain a burst-release artificial bear bile powder nanoparticle formulation. Figure 1 The XRD pattern of the burst-release artificial bear bile powder nanoparticle formulation is shown. The phase retrieval results indicate that the material was successfully prepared.

[0037] This embodiment uses the above method to prepare a burst-release artificial bear bile powder nanoformulation: (1) The cumulative release rate of the main active ingredients in artificial bear bile powder in phosphate buffer solution at pH 7.4 at 37°C for 24 hours was 35%; the cumulative release rate of the main active ingredients in artificial bear bile powder in phosphate buffer solution at pH 5.5 at 37°C for 2 hours was 45.2%; and the cumulative release rate of the main active ingredients in artificial bear bile powder in 24 hours was 90%.

[0038] (2) Its drug loading is 19.7%.

[0039] (3) It is a composite nanoparticle powder with the following physical structure, such as Figure 4 As shown, artificial bear bile powder is loaded onto a porous framework formed by the self-assembly of magnesium ion-doped calcium phosphate nanocrystals. The framework consists of nanoparticle aggregates, a network of interlaced nanosheets / nanose needles, or a flower-like structure of stacked micron-scale sheets.

[0040] (4) Its components include artificial bear bile powder and zinc oxide nanoparticles, and the artificial bear bile powder includes the following components: Tauroursodeoxycholic acid 40-55%; Taurine chenodeoxycholic acid 15-30%; Ursodeoxycholic acid 2-5%; Chenodeoxycholic acid 1-3%; Bile acids 5-10%; The bile acid components simultaneously satisfy the following proportional relationships, specifically: The mass ratio of tauroursodeoxycholic acid (TUDCA) to taurourchedeoxycholic acid (TCDCA) is 2.5:1; and the mass fraction of taurine-conjugated bile acids in total bile acids is greater than 80% and less than or equal to 100%, and the taurine-conjugated bile acids are the sum of tauroursodeoxycholic acid (TUDCA) and taurourchedeoxycholic acid (TCDCA). Total bile acids include free bile acids, glycine-conjugated bile acids, and taurine-conjugated bile acids.

[0041] In summary, among Examples 1-3, the one with the best performance is the burst-release artificial bear bile powder nanoparticle preparation obtained in Example 3, which can be used in the treatment of diseases that require rapid drug release in an acidic microenvironment.

[0042] The technical scope of this invention is not limited to the content described above. Those skilled in the art can make various modifications and variations to the above embodiments without departing from the technical concept of this invention, and all such modifications and variations should fall within the protection scope of this invention.

Claims

1. A method for preparing a burst-release artificial bear bile powder nanoparticle formulation, characterized in that, Includes the following steps: Step S1: Preparation of magnesium ion-doped calcium phosphate nanoparticles Step S1.1: Dissolve calcium salt and magnesium salt in deionized water to obtain a mixed cation solution, wherein the molar ratio of magnesium ions to calcium ions is 0.05:1 to 0.2:1; Step S1.2: Dissolve phosphate in deionized water to obtain a phosphate solution, wherein the molar ratio of phosphate ions to calcium ions is 1.6:1 to 1.8:1; Step S1.3: Under continuous stirring at 100-500 rpm using a magnetic stirrer, the phosphate solution is added dropwise to the mixed cation solution, while the pH of the mixture is adjusted to 9.0-11.

0. The mixture is reacted at 60-80℃ for 8-12 hours to obtain magnesium-doped calcium phosphate precipitate. Step S1.4: Remove impurities from the precipitate and dry it to obtain magnesium ion-doped calcium phosphate nanoparticles. Step S2, Loading with artificial bear bile powder Step S2.1: Disperse the magnesium ion-doped calcium phosphate nanoparticles obtained in step S1.4 in anhydrous ethanol to obtain a nanoparticle dispersion. Step S2.2: Dissolve the artificial bear bile powder in anhydrous ethanol to obtain an ethanol solution or dispersion of the artificial bear bile powder; Step S2.3: Add the ethanol solution or dispersion of the artificial bear bile powder obtained in step S2.2 to the nanoparticle dispersion obtained in step S2.1, and continue stirring and mixing. The mass ratio of the magnesium ion-doped calcium phosphate nanoparticles to the artificial bear bile powder is 4:1 to 6:1 to obtain a mixed suspension. Step S2.4: The mixed suspension obtained in step S2.3 is subjected to ultrasonic-assisted light-protected vibration at a temperature of 25-35℃ for 12-24 hours to complete the drug loading, thereby obtaining a drug-loaded suspension. Step S3: Collection and Purification Step S3.1: Centrifuge the drug-loaded suspension obtained in step S2.4 and collect the precipitate; Step S3.2: Wash the precipitate obtained in step S3.1 with anhydrous ethanol; Step S4: Using freeze-drying, the precipitate obtained in step S3.2 is pre-frozen at -40℃ to -80℃ for 2-12 hours, and then sublimated and dried under a vacuum pressure of 10-30 Pa for 16-48 hours to obtain a burst-release artificial bear bile powder nanoparticle formulation.

2. The method for preparing a burst-release artificial bear bile powder nanoparticle formulation according to claim 1, characterized in that, In step S1.1, the calcium salt and magnesium salt need to be completely dissolved in deionized water by magnetic stirring at a temperature of 15-30°C. The calcium salt is calcium chloride dihydrate and the magnesium salt is magnesium chloride. In step S1.2, the phosphate needs to be completely dissolved in deionized water by magnetic stirring at a temperature of 15-30℃. The phosphate is diammonium hydrogen phosphate. In step S1.3, the pH adjustment process of the mixture is to add alkaline pH adjuster ammonia water dropwise by a metering pump connected to an automatic pH controller, and monitor the pH of the mixture in real time so that the pH is dynamically maintained in the range of 9.0-11.

0.

3. The method for preparing a burst-release artificial bear bile powder nanoparticle formulation according to claim 1, characterized in that, In step S2.1, the process of dispersing magnesium ion-doped calcium phosphate nanoparticles in anhydrous ethanol is specifically as follows: a high shear disperser is used to pre-disperse the nanoparticles at a speed of 3000 rpm for 10 min, and then a high pressure homogenizer is used to cycle and homogenize them 5 times under a pressure of 150 MPa to obtain a uniformly dispersed nanoparticle dispersion. In step S2.2, the particle size requirement for artificial bear bile powder is D90 < 50 µm.

4. The method for preparing a burst-release artificial bear bile powder nanoparticle formulation according to claim 1, characterized in that, Specifically, step S3.1 involves centrifuging the drug-loaded suspension obtained in step S2.4 at 1000-4000 rpm for 10-30 minutes, collecting the precipitate, and repeatedly centrifuging and redispersing the collected precipitate with anhydrous ethanol, washing it 2-5 times, and collecting the precipitate again.

5. A burst-release artificial bear bile powder nanoparticle preparation obtained by the method described in any one of claims 1-4.

6. The burst-release artificial bear bile powder nanoformulation according to claim 5, characterized in that, The cumulative release rate of the main active ingredient in the artificial bear bile powder of the nano-formulation was less than 40% within 24 hours at 37°C in a phosphate buffer solution with pH 7.4; and greater than 40% within 2 hours and greater than 70% within 24 hours in a phosphate buffer solution with pH 5.5 at 37°C.

7. The burst-release artificial bear bile powder nanoformulation according to claim 5, characterized in that, The drug loading of the formulation is 10-20%.

8. The burst-release artificial bear bile powder nanoformulation according to claim 5, characterized in that, The formulation is a composite nanoparticle powder with the following physical structure: the artificial bear bile powder is loaded on a porous framework formed by the self-assembly of magnesium ion-doped calcium phosphate nanocrystals. The framework is composed of nanoparticle aggregates, a network of interlaced nanosheets / nanose needles, or a flower-like structure of stacked micron-scale sheets.

9. The burst-release artificial bear bile powder nanoformulation according to claim 5, characterized in that, It includes artificial bear bile powder and zinc oxide nanoparticles, wherein the artificial bear bile powder comprises the following components: Tauroursodeoxycholic acid 40-55%; Taurine chenodeoxycholic acid 15-30%; Ursodeoxycholic acid 2-5%; Chenodeoxycholic acid 1-3%; Bile acids 5-10%; The bile acid components satisfy one or more of the following proportional relationships, specifically: The mass ratio of tauroursodeoxycholic acid (TUDCA) to taurourcheideoxycholic acid (TCDCA) is (1.5-2.5):

1. b) The mass fraction of taurine-conjugated bile acids in total bile acids is greater than 80% and less than or equal to 100%, and the taurine-conjugated bile acids are the sum of tauroursodeoxycholic acid (TUDCA) and taurourchedeoxycholic acid (TCDCA), and the total bile acids include free bile acids, glycine-conjugated bile acids and taurine-conjugated bile acids.

10. The use of the burst-release artificial bear bile powder nanoformulation according to any one of claims 1-4 in the preparation of a medicament for treating diseases requiring rapid release of the drug in an acidic microenvironment, wherein the disease is selected from hepatobiliary inflammation or inflammatory bowel disease.

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